Evans blue

The following reagents were obtained from the indicated suppliers: Paclitaxel injection (5 mL: 30 mg, the excipients in the paclitaxel injection are polyoxyl 35 castor oil, anhydrous citric acid and anhydrous ethanol, Beijing Union Pharmaceutical Factory, Beijing, CHN), relevant vehicle (polyoxyl 35 castor oil and anhydrous ethanol, 1:1 v/v, Beijing Union Pharmaceutical Factory, Beijing, CHN), compound 48/80 (Sigma-Aldrich, Louis, MO, USA), Evans blue (Sinopharm Chemical Reagent Co., Ltd., Shanghai, CHN), and Cobra venom factor (Shanxi Powerdone Pharmaceutics Co., Shanxi, CHN).
The following antibodies were used in this study: anti-p-MYPT1 (Thr 696) (rabbit polyclonal, 5163, Cell Signaling Technology (CST), MA, USA); anti-MYPT1 (rabbit polyclonal, 2634, CST, MA, USA), anti-p-MLC2 (Thr18/Ser19) (rabbit polyclonal, 3674, CST, MA, USA), anti-MLC2 (rabbit polyclonal, 3672, CST, MA, USA), Anti-RhoA (67B9) (rabbit monoclonal, 2117, CST, MA, USA), anti-ROCK1 (rabbit monoclonal, EP786Y, Abcam, Cambridge, UK), anti-GAPDH (rabbit polyclonal, FL-335, Santa Cruz Biotechnology, CA, USA) and the secondary goat anti-rabbit IgG (H + L) antibody (ZSGB-BIO, Beijing, CHN).

Graphite, Evans blue and Methyl blue were purchased from Sinopharm Chemical Reagent Co., Ltd. Potassium permanganate (KMnO₄, A.R. grade), nitric acid (HNO₃, A.R. grade), hydrochloric acid (HCl, A.R. grade) and hydrogen peroxide (H₂O₂, A.R. grade) were purchased from Yantai Sanhe Chemical Co., Ltd. Direct red 81 (DR81) was purchased from TCI (Shanghai) Development Co., Ltd. Rhodamine B was purchased from Tianjin Dengke Chemical Co., Ltd. 4-Nitrobenzene diazonium tetrafluoroborate (NDT) was purchased from Sigma-Aldrich. Mixed cellulose ester membranes (MCE, 50 mm in diameter), as supporting membranes with a pore size of 0.22 μm, were provided by Shanghai Xingya material works.

ABTS, acrylamide, bisacrylamide, glycine, sodium dodecyl sulfate (SDS), and Tris-base were purchased from Sigma-Aldrich Chem. Co. (United States). DEAE-Sepharose, Q-Sepharose, SP-Sepharose, Superdex 75 HR 10/30 column, and molecular mass standards for FPLC were obtained from GE Healthcare Co. Ltd (Boston, MA, United States). The protein marker for SDS-PAGE was purchased from Biomed SciTech Co., Ltd. (Beijing, China). Synthetic dyes including active red (AR), bromophenol blue (BB), Coomassie brilliant blue G250 (CBB), Evans blue (EB), eriochrome black T (EBT), malachite green (MG), methyl orange (MO), reactive black (RB), reactive brilliant blue (RBB), reactive orange (RO), reactive red (RR), reactive turquoise blue (RTB), reactive yellow (RY), trypan blue (TB), and mediators including acetosyringone (AS), HBT, syringaldehyde (SA), 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO), vanillin (VAL), and violuric acid (VIA) were purchased from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China). Other chemicals used were of analytical reagent grade.